3-hydroxy-5-pregnene-7, 11, 20-triones and esters thereof



United States Patent 3-HYDROXY-5-PREGNENE-7,11,20-TRIONES AND ESTERSTHEREOF Herbert C. Murray, Hickory Corners, and Durey H. Peterson,Kalamazoo Township, Kalamazoo County, Mich., assignors to The UpjohnCompany, Kalamazoo, Mich a corporation of Michigan No Drawing.Application August 26, 1953, Serial No. 376,738

6 Claims. (Cl. 260*39745) u. 5.1% mt,

wherein R is hydrogen or a hydrocarbon-earboxylic acyl radical, areprepared by reacting 3/8,7,lla-trihydroxy- -pregnene-20-one or35,1la-dihydroxy-5-pregnene-7,2Q- dione with an oxidizing agent toproduce 3fi-hydroxy-5- pregnene-7,l1,20-trione which may then 'beesterified. The starting compounds utilized in the process of thepresent invention may be prepared by subjecting pregnenolone(3fi-hydroxy-5-pregnene-20-ohc) to the air genating action or" a cultureof fungus of the order Mucoraies as set forth in our applications, ofwhich the present application is a continuation in-part, Serial No.180,496, filed August 19, 1950, now abandoned, Serial No. 272,944, filedFebruary 23, 1952, issued July 8, 1952, as Patent No. 2,602,769, and'copending Serial No. 296,742, filed July 1, 1952, now abandoned.

'It is an object of this invention to prepare the novel '35--hydroxy-5-pregnene-7;1 1,20-trione and its esters. It is a furtherobject of the present invention to provide a novel process for thepreparation of 3B-hydroxy-5-pregnene 7,11,20-trione. Another object ofthis invention is to provide a novel process for the preparation ofesters of 3 3- hydroxy-5-pregnene-7,11,20-trione. Other objects will beapparent to those skilled in the art to which this invention pertains.

The thermostable compounds of the present invention have exhibitedpharmacological properties and are useful as chemical intermediates.They exhibit anti-hypertensive, anti-estrogenic, anti-progesterone,anti=testoid, antifolliculoid, and anaesthetic activities. The compoundsof the present invention may be converted to the knownll-ketoprogesterone, by hydrogenation with lithium aluminum hydride togive 3-acyloxy-7,11,20-trihydroxyor 3,7,1l,20-tetrahydroxy-S-pregnenewhich is oxidized with manganese dioxide in chloroform to3-acyloxy-1l,20- dihydroxyor 3,1l,20-trihydroxy-5-pregnene-7-one. Thethus-obtained 7lcetone is reduced by the Wolif-Kishner method to give3-acyloxy-l1,20-dihydroxy-5-pregnene or 3,11,20 trihydroxy 5 pregnene.3,11,20-trihydroxy-5- pregnene thus-obtained or prepared bysaponification of 3 acyloxy 11,20-dihydroXy-5-pregnene is oxidized withchromic acid to ll-ketoprogesterone.

in the process of the present invention, the starting steroid selectedfrom the group consisting of 35,7;11a-trihydroxy-S-pregnene-ZO-one and3,6,11t-dihydroxy-5-pregnene-7,20-dione is dissolved in a solvent suchas, for example, glacial acetic acid, and oxidized with a suitableoxidizing agent such as, for example, chromium trioXide, chromic acid,potassium permanganate, N-bromo-succinimide in the presence of an acidbinding agent, and others. Chromium trioxide has been found to be asatisfactory oxidizing agent, converting the starting steroid to desiredproduct in high yield. The oxidizing agent is usually used in proportiongreater than theoretical to completely convert the starting steroid to3B-hydroxy-5pregnene-7,- 11,20-trione, although other proportions arewithin the scope of the present invention. The oxidizing agent isusually added at room temperature and thereafter the reaction mixture isallowed to stand at a temperature between about zero degrees centigradeand the boiling point of the reaction mixture, usually room temperature.The time of reaction is dependent, in part, upon the oxidizing agentemployed and the temperature at which the reaction is carried out. Thesolvent may then be removed by distillation, under vacuum, and thedesired product precipitated by the addition of water whichconcomitantly dissolves any inorganic salts present. The 3fi-hydroxy-5-pregnene-7,11,20-trione thus-produced may then be purified byfiltration and subsequent recrystallization from a suitable solvent suchas, for example, isopropyl alcohol, or chromatographic separationaccording to methods known in the art.

The following examples are illustrative of the process and products ofthe present invention, but are not to be construed as limiting.

Preparation 1 .3 B, 7,] 1 a-trihydr0xy-5-pregnene-20-0ne A medium wasprepared of twenty grams of Edamine enzymatic digest of lactalbumin,three grams of corn steep liquor and fifty grams of technicaldextrosediluted to one liter with tap water and adjusted to a pH of 4.3to 4.5. Eight liters of this sterilized medium was inoculated withRhizopus arrhizus ATCC 11145 and incubated for nineteen hours at atemperature of 28 degrees centigrade using a rate of aeration andstirring such that the oxygen uptake was 6.3 tof7 millimoles per hourper liter of NazSOs according to the method of Cooper, Fernstrom andMiller, Ind. Eng. Chem., 36, 504 (1944). To this medium containing anineteenhour growth of Rhizopus arrhizus was added two grams ofpregnenolone (3,6-hydroxy-S pregnenc ZO-one) in twenty milliliters ofacetone to provide a suspension of the steroid in the culture. After anadditional -hour period of incubation under the same conditions oftemperature and aeration, the beer and mycelium were extracted andconcentrated. The mycelium was filtered, washed twice, each time with avolume of acetone approximately equal to the volume of the mycelium andextracted twice, each time with a volume of methylene chlorideapproximately equal to the volume of the mycelium. The acetone andmethylene chloride extracts including solvent were added to the beerfiltrate. The mixed extracts and beerfiltrate were extractedsuccessively with four liters of methylene chloride and then with threetwo-liter portions of methylene chloride. The combined methylenechloride extracts were washed with two one-tenth by volume portions of atwo percent aqueous solution of sodium bicarbonate and then with twoone-tenth by volume portions of water. After dry ing the methylenechloride with about three to five grams of anhydrous sodium sulfate perliter of solvent and filtering, the solvent was removed by distillationleaving 3.3162 grams of crystalline residue. This residue was dissolvedin 200 milliliters of benzene and chromotographed over grams of alumina(acid washed, dried at degrees centigra de for four hours). The columnwas developed with ZOO-milliliter portions of solvents as indicated inTable I.

Fractions 28 through 31 were combined, dissolved in ten milliliters ofmethanol, and concentrated until crystallization ensued. A few drops ofwater were added, the mixture was cooled, and the crystals wereseparated. Three recrystallizations from methanol, addingwater tofacilitate crystallization, or methanol-carbon tetrachloride yielded 375milligrams of 3fl,7,1 la trihydroxy-5- preg'nene-20-one, melting point247 to 248 and 250 to 255 degrees Centigrade, [di of minu's 41''degr'ees (1.086 in methanol). spectra.

Analysis.Calculated for C21H32042 Found: C, 72.34; H, 9.13.

Preparation 2.-,3fi,1 1 a-dihydroxy-S-pregnene-7,20-dione Example1.--3f3-hydr0xy-5-pregncue-7,1LZO-triane A 28-milligram sample of3,B,1le-dihydroxy-5-pregnenee7,20-dione was dissolved in two millilitersof glacial acetic acid and a solution of 200 milligrams of chromiumtrioxide in a mixture of 0.2 milliliter of water and two milliliters ofglacial acetic acid. The reaction mixture was kept at room temperaturefor sixteen hours whereafter the mixture was diluted with tenmilliliters of methanol, and the volatile components thereafterdistilled under vacuum. The residue was mixed with thirty milliliters ofwater and the whole extracted with three ten-milliliter portions ofether. The combined ether extracts'were washed with two ten-milliliterportions of a five percent sodium bicarbonate solution followed by twoten-milliliter portions of water whereafter the ether layer was driedwith anhydrous sodium sulfate. The ether layer was evaporated leaving acrystalline residue of 20.5 milligrams of Sfl-hydroxy-S-pregnene-Zl1,20-trione which melted at 228 to 229 degrees centigrade after tworecrystallizations from 0.5 milliliter of ethyl acetate to which wasadded .Skellysolve B, petroleum ether, to the point of turbidity.Infrared absorption spectrum confirmed the theoretical structure. Theprodnot had an [M1 of minus 34 degrees (0.935 in chloroform).

Analysis-Calculated for C21H2804! C, 73.23; H, 8.19. Found: C, 73.04; H,8.42.

Example 2.-3fl-hydroxy-5-pregnene-7,11,20-tri0ne A 27.5-milligram sampleof 35,7,11a-t1ihYd10XY-5- pregnene-ZO-one was dissolved in fivemilliliters of glacial acetic acid and a solution of twenty milligramsof chromium trioxide in a mixture of a few drops of water and twomilliliters of glacial acetic acid was then added thereto. The reactionmixture was kept at room temperature for fifteen hours whereafter themixture was diluted with ten milliliters of methanol and the volatilecomponents thereafter distilled under vacuum. The residue was mixed withtwenty milliliters of water and the whole extracted with threetwenty-milliliter portions of ether. The combined ether extracts werewashed with two five-milliliter portions of a five percent sodiumbicarbonate solution followed by three five-milliliter portions of waterwhereafter the ether layer was dried'with anhydrous sodium sulfate. Theether was evaporated leaving a crystalline residue of 14.5 milligrams of3B- hydroxy-5-pregnene-7,l1,20-trione which melted at 227 to 228 degreescentigrade upon recrystallization from an acetone and ether mixture.Infrared absorption spectrum confirmed the identity of the product whichwas identical to the product obtained in Example 1.

Example 3 .-3fi-acetoxy-5 -pregnene-7,1 1 ,ZO-trione A fifty-milligramportion of 3p-hydroxy-5-pregnene- 7,11,20-trione was dissolved in fivemilliliters of acetic anhydride and five milliliters of pyridine. Themixture was maintained at room temperature for 48 hours and then dilutedto 100 milliliters with water. The diluted mixture was extractedsuccessively with eighty-, forty-, and forty-milliliter portions ofether. The combined ether extracts were washed successively with fifteenmilliliters of water, twice with fifteen-milliliter portions of normalhydrochloric acid, fifteen milliliters of water, three times with25-milliliter portions of sodium bicarbonate solution, and three timeswith 25-milliliter portions of water. The washed ether solution wasdried over anhydrous sodium sulfate and evaporated by a stream of air.The residue was taken up in methanol, freed of solvent, and twicereprecipitated from two milliliters of Structure was verified byinfrared acetone by the dropwise addition of ether. Filtration yielded3fl-acetoxy-5-pregnene-7,l1,20-trione. Example 4.3fi-pr0pi0nyl0xy-5-pregnene-7,1 1,20-trione In the same manner as given in Example 2, the3,8- propionyloxy-5-pregnene-7,11,20-trione is prepared using theequivalent proportion of propionic anhydride in place of aceticanhydride.

Example 5 .-3;8-benz0xy-5 -pregnene-7,1 1 ,ZO-trione In the same manneras given in Example 2, the 3 8- benzoxy-5-pregnene-7,l1,20-trione isprepared using the equivalent proportion of propionic anhydride in placeof acetic anhydride. V

Esters of 3fi-hydroxy-5-pregnene-7,11,20-trione are prepared as shownabove or according to various acylation procedures, as by reaction withketene, ketenes of selected acids, selected acids, acid anhydrides, oracid chlorides, in an organic solvent such as pyridine or the like.Representative esters of 3B-hydroxy-5-pregnene- 7,11,20-trionethus-prepared include one to eight carbon atom carboxylic acid acyloxyesters of saturated or unsaturated aliphatic or cycloaliphatic,carboxylic, aryl,

arylalkyl, alkaryl, mono, di or polycarboxylic acids which form estergroups such as, for example, formyloxy, acetoxy, propionyloxy,dimethylacetoxy, trimethylacetoxy, butyryloxy, valeryloxy, hexanoyloxy,heptanoyloxy, octanoyloxy, benzoxy, phenylacetoxy, toluoloxy,naphthoyloxy, cyclopentylformyloxy, B-cyclopentylpropionyloxy,acrylyloxy, cyclohexylformyloxy, the half and diesters of malonic,maleic, succinic, glutaric and adipic acids, and the like. The acids mayalso contain noninterfering substituents, such as mono or poly halo,chloro,

' bromo, hydroxy, methoxy, and the like, if desired.

TABLE I Eluate Fraction Solvent Solids Milligrams 1,2 benzene 195 56 l1ether 7 ether plus 5 percent CECI 8 either plus 10 percent OHOI 32 etherplus 50 percent OHGI; 108 chloroform 590 CHO]; plus 5 percent acetone---37 (in 24 CHO]; plus 10 percent acetone 0110!; plus 50 percent acetone171 399 do 110 acetone plus 5 percent methanol-.. 244 do 125 30,31.acetone plus 10 percent methanol 118 32,33. acetone plus 50 percentmethanoL. 73 ,35 methanol 27 It is to be understood that the inventionis not to be limited to the exact details of operation or exactcompounds shown and described as obvious modifications and equivalentswill be apparent to one skilled in the art and the invention istherefore to be limited only by the scope of the appended claims.

We claim:

l. A compound represented by the formula:

wherein R is selected from the radicals consisting of 6 hydrogen andhydrocarbon-carboxylic acyl having less 4.3fl-acetoxy-5-pregnene-7J1,20-trione.

an nine carbon atoms. 5. 3fi-propionyloxy-5-pregnene-7,11,20-trione. 2.3B-hydroxy-5-pregnene-7,11,20-trione. 6.3fi-benzoxy-5-pregnene-7,11,20-trione. 3.SB-acyloxy-S-pregnene-Zl1,20-trione wherein acyloxy is ahydrocarbon-carboxylic radical containing less 5 No references cited.

than nine carbon atoms.

1. A COMPOUND REPRESENTED BY THE FORMULA: